pMIB/V5-His is a 3.6 kb vector designed for constitutive expression and secretion of recombinant proteins from Sf9, Sf21, and High Five insect cells. Secretion of proteins into serum-free medium simplifies purification, making it easier to harvest protein from cultured cells. pMIB/V5-His has several features to facilitate expression of recombinant proteins in insect cells including:
• The OpIE2 promoter for constitutive expression
• The honeybee melittin (HBM) secretion signal to allow secreted expression of your protein
• The Blasticidin resistance gene for rapid selection of stably transfected cell lines in two weeks
• C-terminal V5-epitope and polyhistidine tag for detection with Invitrogens Anti-V5 antibody and purification with nickel-chelating resin
引用及参考文献
Antibodies against a secreted protein from hookworm larvae reduce the intensity of hookworm infection in humans and vaccinated laboratory animals.
Bethony J, Loukas A, Smout M, Brooker S, Mendez S, Plieskatt J, Goud G, Bottazzi ME, Zhan B, Wang Y, Williamson A, Lustigman S, Correa-Oliveira R, Xiao S, Hotez PJ, FASEB J (2005) 19:1743-1745
Product usage: ASP-2 was fused in-frame into pMIB-V5/HisA using the SphI and XbaI restriction sites. Protein was expressed in Spodpotera frugiperda Sf9 insect cells.
CATGATGATAAACAATGTATGGTGCTAATGTTGCTTCAACAACAATTCTGTTGAACTGTGTTTTCATGTT
TGCCAACAAGCACCTTTATACTCGGTGGCCTCCCCACCACCAACTTTTTTGCACTGCAAAAAAACACGCT
TTTGCACGCGGGCCCATACATAGTACAAACTCTACGTTTCGTAGACTATTTTACATAAATAGTCTACACC
GTTGTATACGCTCCAAATACACTACCACACATTGAACCTTTTTGCAGTGCAAAAAAGTACGTGTCGGCAG
TCACGTAGGCCGGCCTTATCGGGTCGCGTCCTGTCACGTACGAATCACATTATCGGACCGGACGAGTGTT
GTCTTATCGTGACAGGACGCCAGCTTCCTGTGTTGCTAACCGCAGCCGGACGCAACTCCTTATCGGAACA
GGACGCGCCTCCATATCAGCCGCGCGTTATCTCATGCGCGTGACCGGACACGAGGCGCCCGTCCCGCTTA
TCGCGCCTATAAATACAGCCCGCAACGATCTGGTAAACACAGTTGAACAGCATCTGTTCGAATTTAAAGC
TACCATGAAATTCTTAGTCAACGTTGCCCTTGTTTTTATGGTCGTATACATTTCTTACATCTATGCCGGC
ATGCTAAGCTTGGTACCGAGCTCGGATCCACTAGTCCAGTGTGGTGGAATTCTGCAGATATCCAGCACAG
TGGCGGCCGCTCGAGTCTAGAGGGCCCTTCGAAGGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATT
CTACGCGTACCGGTCATCATCACCATCACCATTGAGTTTATCTGACTAAATCTTAGTTTGTATTGTCATG
TTTTAATACAATATGTTATGTTTAAATATGTTTTTAATAAATTTTATAAAATAATTTCAACTTTTATTGT
AACAACATTGTCCATTTACACACTCCTTTCAAGCGCGTGGGATCGATGCTCACTCAAAGGCGGTAATACG
GTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAAC
CGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGAC
GCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCT
CGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTG
GCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTG
TGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGT
AAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGT
GCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTC
TGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAG
CGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATC
TTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGCGAAACACGC
ACGGCGCGCGCACGCAGCTTAGCACAAACGCGTCGTTGCACGCGCCCACCGCTAACCGCAGGCCAATCGG
TCGGCCGGCCTCATATCCGCTCACCAGCCGCGTCCTATCGGGCGCGGCTTCCGCGCCCATTTTGAATAAA
TAAACGATAACGCCGTTGGTGGCGTGAGGCATGTAAAAGGTTACATCATTATCTTGTTCGCCATCCGGTT
GGTATAAATAGACGTTCATGTTGGTTTTTGTTTCAGTTGCAAGTTGGCTGCGGCGCGCGCAGCACCTTTG
CCGGGATCTGCCGGGCTGCAGCACGTGTTGACAATTAATCATCGGCATAGTATATCGGCATAGTATAATA
CGACAAGGTGAGGAACTAAACCATGGCCAAGCCTTTGTCTCAAGAAGAATCCACCCTCATTGAAAGAGCA
ACGGCTACAATCAACAGCATCCCCATCTCTGAAGACTACAGCGTCGCCAGCGCAGCTCTCTCTAGCGACG
GCCGCATCTTCACTGGTGTCAATGTATATCATTTTACTGGGGGACCTTGCGCAGAACTCGTGGTGCTGGG
CACTGCTGCTGCTGCGGCAGCTGGCAACCTGACTTGTATCGTCGCGATCGGAAATGAGAACAGGGGCATC
TTGAGCCCCTGCGGACGGTGCCGACAGGTTCTTCTCGATCTGCATCCTGGGATCAAAGCCATAGTGAAGG
ACAGTGATGGACAGCCGACGGCAGTTGGGATTCGTGAATTGCTGCCCTCTGGTTATGTGTGGGAGGGCTA
AGCACTTCGTGGCCGAGGAGCAGGACTGACACGTCCCGGGAGATCTGCATGTCTACTAAACTCACAAATT
AGAGCTTCAATTTAATTATATCAGTTATTACCCATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCG
TGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAA
GTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGA
TCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGC
GGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTG
GTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTG
CCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAAC
CGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCC
ATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTG
GCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACC
ACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCT
CGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGA
GTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTA
ACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATC
TAGGTGAAGATCCTTTTTGATAATCT